直接竞争酶联免疫法测定食品中的丙烯酰胺含量  

Direct Competitive Enzyme-Linked Immunosorbent Assay for Detection of Acrylamide in Food Samples

作　　者：吴璟[1] 罗林[1] 肖治理[1] 杨金易[1] 孙远明[1] 雷红涛[1] 沈玉栋[1] 王弘[1] 徐振林[1] 

机构地区：[1]华南农业大学食品学院/广东省食品质量安全重点实验室,广州510642

出　　处：《分析化学》2014年第8期1149-1154,共6页Chinese Journal of Analytical Chemistry

基　　金：973课题(2012CB720803);广东省高等学校学科与专业建设专项资金(2013KJCX0027)项目资助~~

摘　　要：丙烯酰胺由于分子量小、结构简单,制备其特异性抗体难以实现。本研究将丙烯酰胺与对巯基苯乙酸衍生合成半抗原,偶联载体蛋白并免疫动物制备针对丙烯酰胺衍生物的特异性抗体,并进行辣根过氧化物酶标记,进而建立通过检测丙烯酰胺衍生物实现对丙烯酰胺定量分析的直接竞争酶联免疫分析方法。本方法对丙烯酰胺的检出限为3.0μg/L,线性范围为9.2~195μg/L,对饼干、薯片及咖啡样品中丙烯酰胺的平均添加回收率为83.6%~112.7%,结果与标准检测方法 HPLC-MS/MS符合。本方法可用于食品样品中丙烯酰胺的快速检测。Due to the low molecular weight and simple structure, the production of specific antibodies against acrylamide is unavailable. In this study, a novel hapten was synthesized through the derivatization of acrylamide and 4-mercaptophenylacetic acid. The hapten was then coupled to carrier protein and used to immunize New Zealand rabbits. Polyclonal antibody which showed specific binding to the acrylamide derivative （ hapten） was obtained. The antibody was labeled with horseradish peroxidase （ HRP） and used to develop a direct competitive enzyme-linked immunosorbent assay （ dc-ELISA） . The dc-ELISA was used to determine the content of acrylamide derivative, and then transferred to the content of acrylamide. The assay showed an IC50 value of 45. 49 μg/L, a limit of detection of 3. 0 μg/L and the linear range of 9. 2-195 μg/L for acrylamide. The recovery of acrylamide from spiked food sample was determined ranging from 83 . 6% to 112 . 7%. Good correlations between the results of dc-ELISA and standard HPLC-MS/MS were obtained. The proposed dc-ELISA is suitable for the determination of acrylamide in food samples.

关 键 词：丙烯酰胺 多克隆抗体 直接竞争酶联免疫分析 衍生 

分 类 号：TS207.3[轻工技术与工程—食品科学]